Abstract
Since the core protein of mucin in the digesta of the stomach and small intestine, which is less affected by bacteria, remains intact, mucin content can be measured by enzyme-linked immunosorbent assay (ELISA). However, the mucin core protein in bacteria-rich colon digesta and feces is partially hydrolyzed by bacterial enzymes and not fully recognized by mucin antibodies, so mucin cannot be accurately quantified by ELISA. This method quantifies the glycan content linked to the mucin core protein and expresses mucin content in the colon digesta and feces as the equivalent of O-linked oligosaccharide chain. Although mucin glycans are also hydrolyzed by colonic bacteria, this method is a more accurate and simple way to measure mucin content in the digesta of the large intestine and feces than the ELISA method.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Hansson GC (2020) Mucins and the Microbiome. Annu Rev Biochem 89:769–793. https://doi.org/10.1146/annurev-biochem-011520-105053
Hino S, Mizushima T, Kaneko K, Kawai E, Kondo T, Genda T, Yamada T, Hase K, Nishimura N, Morita T (2020) Mucin-Derived O-Glycans Act as Endogenous Fiber and Sustain Mucosal Immune Homeostasis via Short-Chain Fatty Acid Production in Rat Cecum. J Nutr 150(10):2656–2665. https://doi.org/10.1093/jn/nxaa097
Tanabe H, Sugiyama K, Matsuda T, Kiriyama S, Morita T (2005) Small intestinal mucins are secreted in proportion to the settling volume in water of dietary indigestible components in rats. J Nutr 135(10):2431–2437. https://doi.org/10.1093/jn/135.10.2431
Crowther RS, Wetmore RF (1987) Fluorometric assay of O-linked glycoproteins by reaction with 2-cyanoacetamide. Anal Biochem 163(1):170–174. https://doi.org/10.1016/0003-2697(87)90108-4
Honda S, Matsuda Y, Takahashi M, Kakehi K, Ganno S (1980) Fluorimetric determination of reducing carbohydrates with 2-cyanoacetamide and application to automated analysis of carbohydrates as borate complexes. Anal Chem 52(7):1079–1082
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Nishimura, N., Tanabe, H., Morita, T. (2024). Extraction of Mucin from Rodent Feces and Determination of O-Linked Oligosaccharide Chain Equivalent Derived from Fecal Mucin. In: Kameyama, A. (eds) Mucins. Methods in Molecular Biology, vol 2763. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3670-1_2
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3670-1_2
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3669-5
Online ISBN: 978-1-0716-3670-1
eBook Packages: Springer Protocols